Fluid sample preparation systems and methods

ABSTRACT

Sample application systems can include an extraction mechanism to remove a sample from sample containers, a sample vessel disposed on a deployment mechanism, where the deployment mechanism is arranged to move the sample vessel to receive a sample, an extraction mechanism washing station to wash the extraction mechanism, a sample applicator to remove a portion of the sample in the sample vessel and apply it onto a sample carrier, where the deployment mechanism can move the sample vessel to a sample application position, a sample vessel washing station to wash the sample vessel, where the deployment mechanism can move the sample vessel to the sample vessel washing station, a sample applicator washing station to wash the sample applicator after the sample has been dispensed onto the sample carrier, and a fluid control system to control flow of a fluid provided to the extraction mechanism and the sample applicator.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of, and claims priority to,U.S. patent application Ser. No. 13/548,773, filed on Jul. 13, 2012,which claims priority to U.S. Provisional Patent Application No.61/510,700, filed on Jul. 22, 2011, the entire contents of each of whichare incorporated by reference herein.

TECHNICAL FIELD

This disclosure relates to the inspection of biological fluid samples,and more particularly to applying biological fluid samples to a surface,such as a sample carrier, e.g., a slide.

BACKGROUND

Systems, such as manufacturing systems or systems for analyzing samples,e.g., fluid samples, tissue samples, food samples, chemical samples,environmental samples, etc., can be used to analyze samples of differentorigins (e.g., body fluids from different patients or environmentalsamples from different regions). To permit automatic or semi-automaticoperation of such systems (e.g., to minimize human interaction),electromechanical systems can be implemented to selectively providesamples to the systems for processing.

SUMMARY

Systems, such as manufacturing or testing systems, e.g., blood analysissystems, having multiple stations, e.g., processing, monitoring, oranalysis stations, can be simplified by creating a sample preparationmechanism that can automatically or semi-automatically remove samples,e.g., fluid samples, tissue samples, food samples, chemical samples, orenvironmental samples, from sample containers and provide the samples tothe manufacturing or testing systems while preventingcross-contamination of samples. In some implementations, the samplepreparation mechanisms can operate via one or multiple fluid handlingaspiration needles and a translating sample vessel that translatesbetween a sample extraction position and a sample application position.

In one implementation, the present disclosure relates to sampleapplication systems that include an extraction mechanism configured toremove a sample from a sample container (e.g., a test tube having acap), a sample vessel disposed on a deployment mechanism, wherein thedeployment mechanism is arranged to move the sample vessel into anextraction position in which the extraction mechanism can dispense asample into the sample vessel, an extraction mechanism washing stationarranged to wash the extraction mechanism after the extraction mechanismhas dispensed the sample into the sample vessel, a sample applicatorarranged to remove a portion of the sample in the sample vessel andapply the portion of the sample onto a sample carrier, wherein thedeployment mechanism is arranged to move the sample vessel into a sampleapplication position in which the sample applicator can remove theportion of the sample in the sample vessel, a sample vessel washingstation arranged to wash the sample vessel after the sample applicatorhas removed the portion of the sample, wherein the deployment mechanismis arranged to move the sample vessel into a position in which thesample vessel washing station can wash the sample vessel, a sampleapplicator washing station arranged to wash the sample applicator afterthe sample applicator has dispensed the portion of the sample onto thesample carrier; and a fluid control system to control flow of a fluidprovided to the extraction mechanism and the sample applicator.

Various implementations and embodiments of the sample applicationsystems can include any one or more of the following features,individually or in combination. The extraction mechanism can include aconduit to penetrate a cap on a test tube. The deployment mechanism caninclude a leadscrew and sliding mechanism. The sample applicator caninclude a conduit to dispense the sample. The washing stations can eachinclude a vessel having a rounded bottom to direct a fluid flow from aconduit inserted into the vessel to the outer surface of the conduit.The fluid control system can include a fluid reservoir, a fluid pump,and a controller to operate the fluid control system.

The sample application systems can further include an informationreading device, wherein the sample container includes machine-readableinformation (e.g., a barcode or a radio-frequency identification tag).The sample application systems can further include a sample modificationsystem, wherein the sample modification system can include a samplediluent system.

In another implementation, the present disclosure relates to methods ofhandling a sample. The methods include receiving a sample containercontaining a volume of a sample, removing a sample from the samplecontainer using an extraction device, dispensing the sample into asample vessel with the extraction device, washing the extraction deviceby dispensing a fluid through the extraction device, moving the samplevessel containing the sample to a sample application position, removinga portion of the sample from the sample vessel using a sampleapplicator, dispensing the sample portion from the sample applicatoronto a sample carrier, rinsing the sample applicator by dispensing fluidthrough the sample applicator, and washing the sample vessel to removeany residual sample.

Implementations and embodiments of the methods described herein caninclude any one or more of the following features, individually or incombination. The extraction devices can be operated by a fluid system,wherein the fluid of the fluid system is separated from the sample by anair pocket within the extraction device. The methods can further includemodifying the sample dispensed into the sample vessel, wherein modifyingthe sample can include adding a diluent fluid to the sample in thesample vessel. Removing a sample from the sample container can includeinserting a needle through a cap attached to a test tube.

The extraction devices and the sample applicators can retain the samplewhen the fluid control system generates a vacuum in the extractiondevice and the sample applicator. Rinsing the extraction device andrinsing the sample applicator can include inserting a portion of theextraction device and a portion of the sample applicator into respectivereceptacles having curved bottoms, such that fluid dispensed from theextraction device and the sample applicator is directed along an outersurface of the extraction device and the sample applicator. Dispensingthe sample portion from the sample applicator can include dispensing thesample portion onto a glass slide. The sample can include a body fluid(e.g., blood).

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and not intendedto be limiting.

Other features and advantages will be apparent from the followingdetailed description, and from the claims.

DESCRIPTION OF DRAWINGS

FIG. 1 is a perspective view of a sample preparation system.

FIG. 2 is a perspective view of an analysis system for analyzing samplessuch as fluids, e.g., body fluids.

FIG. 3 is a flow chart of one embodiment of a sample preparation system.

FIG. 4 is a perspective view of an example of a sample preparationsystem.

FIG. 5A is a perspective view of a tube gripping device transporting atest tube to an inverting mechanism.

FIG. 5B is a perspective view of a cap detection device of a tubegripping device.

FIG. 5C is a bottom view of a gear system of the tube gripping device ofFIG. 5B.

FIG. 5D is a cross sectional view of a rotating assembly of the tubegripping device of FIG. 5A.

FIG. 5E is a perspective view of a test tube priority drawer.

FIG. 5F is a perspective view of a test tube stop securing a small testtube.

FIG. 5G is a perspective view of the test tube stop of FIG. 5F in aclosed position securing a large test tube.

FIG. 5H is a side view of an open mode port aspirator in a stowedposition.

FIG. 5I is a side view of the open mode port aspirator of FIG. 5J in adeployed position.

FIG. 5J is a perspective view of the open mode port aspirator of FIG. 5Jin a deployed position.

FIG. 6 is a perspective view of an inverting mechanism rotating a testtube to a position for fluid extraction.

FIG. 7 is a perspective view of an extraction needle extracting a samplefrom a test tube.

FIG. 8 is a perspective view of the extraction needle of FIG. 7 rotatingto provide the sample to a sample vessel.

FIG. 9 is a perspective view of the sample vessel of FIG. 8 translatingto a diluent position under a diluent needle.

FIG. 10 is a perspective view of the extraction needle of FIG. 7 beinginserted into a wash cup.

FIG. 11 is a perspective view of the sample vessel of FIG. 8 translatingto a sample application position under an application conduit.

FIG. 12 is a perspective view of the application conduit of FIG. 11applying a sample to a sample carrier.

FIG. 13 is a perspective view of the sample vessel of FIG. 8 translatingto engage a sample vessel wash system.

FIG. 14 is a perspective view of the sample vessel of FIG. 8 translatingback to the sample extraction position under the extraction needle.

FIG. 15 is a perspective view of the application conduit inserted in awash cup.

FIG. 16A is a schematic diagram of an open mode port aspirator in adeployed position.

FIG. 16B is a perspective view of a pivot block for rotating anaspirator probe to a deployed position.

FIG. 16C is a perspective view of an automated blood analyzer.

FIG. 17 is a cross-sectional view of a sample vessel.

Like reference symbols in the various drawings indicate like elements.

DETAILED DESCRIPTION

For testing biological fluids and tissue, such as blood and other bodilyfluids, the new systems and methods described herein can be used toreceive the fluid (e.g., blood), specimen, or sample from a specimencontainer and apply it to a surface, such as a sample carrier, e.g., aglass or plastic slide, e.g., a microscope slide or cover slip, forprocessing and/or inspection at other locations or in other moduleswithin a larger system (e.g., an analysis system).

Sample Preparation Systems

FIG. 1 shows a sample preparation system 21 for preparing a fluid sampleand applying the fluid sample to a surface such as a sample carrier,e.g., a slide. The sample preparation system 21 can include a samplecontainer carrier 23, an extraction needle 25, a fluid reservoir 26 a, afluid pump 26 b, a fluid system controller 26 c, one or more wash cups27, a modification system 29 such as a diluent system, a sampleapplicator 31, a sample vessel movement mechanism 33 to move a samplevessel 35 to the other components in the sample preparation system 21,and a sample vessel wash system 37.

One or more sample containers 39 can be provided to the samplepreparation system 21 to contain and separate different samples, such assamples from different origins (e.g., body fluid samples from differentpatients or environmental samples from different locations). The samplecontainers 39 can include small cups or cylinders (e.g., test tubes).The sample containers 39 can be sealed to contain the sample when thesample is transported by the sample container carrier 23. In someimplementations, sample containers 39 can include caps 41 (e.g., plug,stopper, cover, lid, or similar device, e.g., made of rubber, silicone,or plastic) to seal the sample containers.

The sample containers 39 can include sample information 43 regarding thesample contained therein. Sample information 43 can include things suchas sample origin (e.g., name of patient that provided the sample orgeographical location where the sample was obtained), type of sample(e.g., type of body fluid, type of environmental sample), time and datethat the sample was obtained from its natural environment (e.g., whenbiological fluid was obtained from a body, when an environmental samplewas removed from the environment). In some implementations, the sampleinformation 43 can be in the form of a barcode or radio-frequencyidentification (“RFID”) tag, or other machine-readable format, forsimplified reading and processing by a control unit, e.g., with abarcode reader.

The sample containers 39 can be provided to the sample preparationsystem 21 in a sample container magazine 45. The sample containermagazine 45 can be a device used to transport multiple sample containers39 to and from the sample preparation system 21. The sample containermagazine 45 has multiple openings or apertures to temporarily supportthe sample containers 39 such that the sample containers do not spill orlose the sample during transport (e.g., test tube sample containers aresupported and held upright).

The sample container carrier 23 is used to remove sample containers 39from the sample container magazine 45 so a sample aliquot or portion canbe removed from the sample container 39. The sample container carrier 23can operate in various ways to remove the sample container 39 from thesample container magazine 45. In some implementations, the samplecontainer carrier 23 can be in the form of an articulating roboticdevice that can move to a location of a particular sample container 39(e.g., where the particular sample container 39 is positioned in thesample container magazine 45), grip the particular sample container 39,and lift sample container 39 from the sample container magazine 45 highenough to clear other sample containers 39 held by and from protrudingfrom the sample container magazine 45.

Alternatively, in other implementations, instead of having a roboticdevice that can move laterally to select a particular sample container39 and also have the ability to lift the sample container 39 from thesample container magazine 45, the sample container carrier 23 caninclude a device to push sample containers 39 from the sample containermagazine 45 by the bottom of the sample containers 39. Once the samplecontainer 39 is pushed from sample container magazine 45 high enough toclear the other sample containers 39 held in the sample containermagazine 45, a robotic device can grip the sample container 39 to removeit from the sample container magazine 45. After the sample container 39is removed from the sample container magazine 45, it is transported toextraction needle 25.

In addition to transporting the sample container 39 from the samplecontainer magazine 45, the sample container carrier 23 can also be usedto prepare the sample for extraction. In some implementations, thesample container carrier 23 can include an inverting mechanism 47 suchthat the sample container 39 can be moved (e.g., rotated) to agitate thesample (e.g., re-suspend blood cells in a blood sample or to mix anon-homogeneous sample). For example, inverting mechanism 47 can rotatesample container 39 multiple times (e.g., 2, 3, 4, 5, or 10 or more)from an upright position to another position, e.g., to 90° or 180° fromthe upright position, so that the sample container is turned upsidedown, e.g., to re-suspend or mix a sample before aspiration. Invertingmechanism 47 can rotate sample containers through other degrees ofinversion (e.g., 45°, 270°, or 360°). In some implementations, theinverting mechanism 47 rotates the sample container 180° so that thesample container is upside down (e.g., the cap 41 is pointed downwardtoward an extraction needle 25) to ensure that the extraction needlealways contacts the sample within the sample container (rather thancontacting air if the needle were inserted from the top into a containerthat is not completely filled with sample). After the sample has beenremoved, the sample container 39 is either returned to the magazine 45(e.g., to the same location from which it was removed, or to adifferent, empty location in the magazine from which it was removed or adifferent magazine) or discarded.

Although the sample container carrier 23 has been described as a deviceused to remove a sample container 39 from a sample container magazine45, in other implementations, sample containers 39 are not provided tothe sample preparation system 21 using a sample container magazine 45.In such implementations a sample container 39 may be manually providedto the sample container carrier 23 by an operator.

The extraction needle 25 is a device that can be inserted to penetrate acap 41 of the sample container 39 to extract a sample portion from thesample container 39. In some implementations, the extraction needle 25and the cap 41 are designed such that after the extraction needle 25 isremoved from the cap 41, the cap 41 automatically seals the hole made bythe extraction needle 25, e.g., by an elastic or resilient nature of thematerial used to make cap 41. To remove fluid samples withoutcross-contaminating the extraction needle 25 or other samples, variousmaterial handling methods can be used. In some implementations, toextract and handle a fluid sample using the extraction needle 25, theextraction needle 25 is connected to a pneumatic or hydraulic system,e.g., a fluid system, such as a buffer fluid system, using tubing, e.g.,to contain the fluid, e.g., buffer fluid. Buffer fluids for the fluidsystem that can be used to operate the extraction needle 25 and othercomponents (e.g., sample applicator 21) can be contained in the fluidreservoir 26 a and can be distributed to the components using the fluidpump 26 b and the fluid system controller 26 c. Motion of the bufferfluid (or simply air or other inert gas, such as nitrogen) within thetubing can be controlled by the fluid pump 26 b to generate a vacuum orpressure to withdraw, control, and expel a fluid sample from the tip ofthe extraction needle 25.

The extraction needle 25 and the associated tubing have small innerdiameters such that the gas or liquid, e.g., buffer fluid, containedtherein can be suspended in the tubing without leaking. During operationof the system, the tubing, and in some cases a portion of the extractionneedle 25, is filled with the buffer fluid that can be provided by thefluid reservoir 26 a and fluid pump 26 b. The remaining portion of theextraction needle 25 (e.g., the portion open to the surroundingenvironment not containing the buffer fluid) can contain an air gap sothat when the buffer fluid in the extraction needle 25 or the connectedbuffer fluid tubing applies a vacuum within the extraction needle, thevacuum can be used to extract a sample fluid, such that the air gapremains between the fluid, e.g., buffer fluid, and the sample fluid,e.g. blood aspirated from a sample tube. Since the tubing and the amountof fluid contained in the tubing is relatively small, the air gap is notexpected to dissolve or otherwise become entrapped in either the bufferfluid or the sample fluid.

In some implementations in which the sample container 39 is rotated 180°from the upright position (e.g., to invert the sample container 39) sothat the sample fluid is extracted from the end of the sample container39 that is facing downward (e.g., the end of the sample container 39having the cap 41, facing downward), the extraction needle 25 caninclude a mechanism to rotate the extraction needle 25 as well as toarticulate the extraction needle 25 upward, toward and through cap 41and into the sample container 39 to extract the sample and downward,away from the sample container 39 to remove extraction needle 25 fromthe sample container 39. Articulation and rotation of the extractionneedle 25 can be achieved by various devices such as electromechanicaldevices (e.g., servos, slide mechanisms, and/or electric motors) and/ormechanical devices (e.g., cams, gears, and/or leadscrews). In someimplementations, the full range of motion of the extraction needle 25(e.g., translating the extraction needle 25 into a sample container 39,removing the extraction needle 25 from the sample container 39, androtating the extraction needle 25) can be achieved using just one motioninput (e.g., one electric motor) in combination with other mechanicaldevices (e.g., one electric motor combined with a leadscrew and camdevices). In other implementations, multiple electromechanical devicescan be used.

The wash cups 27 are devices used to clean various fluid aspiratingand/or dispensing members (e.g., the extraction needle 25 and the sampleapplicator 31) after the members have handled a sample (e.g., after asample has been extracted and dispensed).

In some implementations, the wash cups 27 utilize the same buffer fluid(e.g., a combined buffer and wash fluid composition, as furtherdescribed below) contained in the fluid reservoir 26 a and provided bythe fluid pumps 26 b used in the hydraulic vacuum system to clean themembers (e.g., the extraction needle 25 and/or sample applicator 31). Insuch implementations, the wash cup 27 can have an inner basin 27 a andan outer basin 27 b with a fluid output device 27 c (e.g., a drain orsuction device), and operate by dispensing a portion of the buffer fluidfrom the member such that the buffer fluid flushes any sample remnantsfrom the inner surface of the member. The inner basin 27 a can bedesigned such that it directs the buffer fluid exiting the member toflow along the outer surface of the needle before flowing into the outerbasin 27 b to drain away from the wash cup 27. By directing the bufferfluid in one continuous flow path (e.g., out of the needle, then alongthe outer surface of the needle, and then to the outer basin 27 b anddrain), the possibility of backflow of a buffer fluid contaminated witha sample is reduced. In such implementations, after buffer fluid hasbeen dispensed into the wash cup 27 and the member is removed from thewash cup 27, the buffer fluid system can withdraw remaining buffer fluidback into the member and connected tubing to create an air pocket or gapso that the member can properly handle the next sample.

In other implementations, a wash cup 27 can be in the form of the cup orvessel having a nozzle to provide a wash fluid solution and/or dryingnozzles to remove any residual cleaning solution from the needles orconduits.

The sample vessel 35 is a vessel, such as a cup used to contain andcarry a fluid sample portion to multiple locations during samplepreparation. In some implementations, the sample vessel 35 can serve asa mixing vessel when the fluid sample portion is modified (e.g.,diluted, buffered, or stained). Since the sample vessel 35 willtypically contain a large number of different samples, such as samplesfrom different origins (e.g., blood from different patients) ordifferent types of samples (e.g., different types of body fluids) itshould be formed from a material that is smooth and non-porous toprevent absorption of the sample into the material. The material shouldalso be chosen such that the sample vessel 35 will be inert and tend torepel liquid (e.g., reduce wetting) so that sample fluids will collectat the bottom of the sample vessel 35 more easily for removal from thesample vessel 35. Such materials can include various types of plastics(e.g., Teflon®, Delrin®, or Noryl®), glasses, or some metal materials,and in some cases the sample vessel material can be polished to increasethe sample vessel's ability to repel residue fluids.

In some implementations, the sample preparation system 21 can include asample vessel wash system 37 to clean the sample vessel 35 and to removeany residual fluid sample remnants. The sample vessel wash system 37 caninclude a nozzle to provide a wash fluid to the sample vessel 35 and asuction head to remove the rinse fluid containing any residual fluidsample.

Alternatively, in some implementations, the sample vessel 35 can includean outlet (e.g., a drain) built into the sample vessel 35 so thatresidual fluid samples, or wash fluid is removed from the sample vessel35 using the drain system.

The sample vessel movement mechanism 33 is a device to move the samplevessel 35 to the various components in the sample preparation system 21to allow for automated or semi-automated treatment of samples (e.g.,minimize human interaction) during sample preparation. In someimplementations, the sample vessel movement mechanism 33 can include atrack 49 on which the sample vessel 35 moves (e.g., slides) and atranslating mechanism 51 (e.g., an electric motor connected to aleadscrew or an actuator) to move the sample vessel 35 to the variouscomponents (e.g., the extraction needle 25, the modification system 29,and/or the sample applicator 31) positioned along the track 49 such thatthe sample vessel 35 can stop at several locations along the track 49.In some implementations, the track 49 and translating mechanism 51 canbe one component having a track 49 and a translating device (e.g., apneumatic linear actuator, an electromechanical linear actuator, or anindexing table) used to move the sample vessel 35 along the track 49.Alternatively, in some implementations, the sample vessel 35 can remainstationary and the various components can move (e.g., the modificationsystem 29 and the sample applicator 31 could be mounted to an indexingtable that rotates to modify and withdraw a fluid held in a stationarysample vessel 35).

The modification system 29 can include various systems to prepare asample for use in an analysis system such as a diluent system, astaining system, and/or an anti-coagulation system. Generally, themodification system 29 includes a device to provide a modifyingsubstance (e.g., a metered amount of diluent such as saline, purifiedwater, or protein solutions) to the sample contained in the samplevessel 35. In some implementations, the modification device can includea modification conduit 53 (e.g., a syringe or a pipette) connected to afluid reservoir to provide the modifying substance (e.g., diluent).

The sample applicator 31 is a device used to remove a sample (e.g., asample prepared with a modifying substance or an unmodified sample) fromthe sample vessel 35 and provide the sample to a surface, such as asample carrier (e.g., a glass slide) of an analysis system (e.g., bodyfluid analysis system). The sample applicator 31 can include anapplication conduit 57 (e.g., a portion of tubing, a needle, a syringetip, and/or a pipette) to withdraw and handle a fluid sample. The sampleapplicator 31 can also include a buffer fluid system connected to theapplication conduit 57, similar to the extraction needle 25, so that thesample applicator 31 can use fluid pumps to withdraw a sample from thesample vessel 35 and dispense the sample onto the sample carrier. Toreach a sample vessel 35, the sample carrier of the analysis system, anda wash cup 27, the sample applicator 31 can include a sample applicatortranslating device 32 to move the sample applicator 31 in differentdirections to the various positions (e.g., multiple axes of motion).

Buffer and Wash Fluid Compositions

Buffer and wash fluids, e.g., combined buffer and wash fluidcompositions, can be used with the sample preparations systems describedherein. A combined buffer and wash solution can be used to operate theextraction needle 25 and sample applicator 21, as well as to clean andflush a sample preparation system. As a wash solution, the compositionscan reduce cross-contamination between biological specimens. In someembodiments, the buffer and wash solution can have reduced or noprecipitation and can be stable over a period of time (e.g., more thantwo weeks, more than one month, more than 6 months, more than one year,more than 1.5 years, or more than two years). Generally, the buffer andwash solution can be an aqueous solution. The solvent can includedistilled water or deionized water. The solution can include a bufferingagent. Examples of buffering agents include HEPES buffer (e.g., HEPESsodium salt and/or HEPES free acid), bis-tris buffer, phosphate, MES,Tris, and organic buffers having a pH between 5 and 8. The buffer andwash solution can include from approximately 0.5 mM (e.g., 25 mM, 50 mM,100 mM, 150 mM, or 200 mM) to approximately 250 mM (e.g., 200 mM, 150mM, 100 mM, 50 mM, or 25 mM) of a buffering agent. For example, thesolution can include approximately 1.0 mM HEPES, which can decrease thelikelihood of pH change due to formation of carbonic acid in an aqueoussolution.

The buffer and wash solution can include one or more antimicrobialagents to inhibit the growth of microorganisms and increase the shelflife of the solution. The antimicrobial agents can be or includebenzalkonium chloride, 5-chloro-2-methyl-4-isothiazolin-3-one,2-methyl-4-isothiazolin-3-one, such as ProClin® (e.g., ProClin 300®),polyamino carboxylic acids (e.g., ethylenediaminetetraacetic acid,disodium ethylenediaminetetraacetic acid, and calcium disodiumethylenediaminetetraacetic acid), azides, thimerosols, merthiolates,and/or antibiotics. In some embodiments, the antimicrobial agentincludes 5-chloro-2-methyl-4-isothiazolin-3-one and2-methyl-4-isothiazolin-3-one. For example, the antimicrobial agent canbe ProClin 300®, available from Sigma-Aldrich. The antimicrobial agentcan be present at a concentration of approximately one part in 1,000 toone part in 10,000 (e.g., one part in 2,000, one part in 4,000, one partin 6,000, or one part in 8,000). In some embodiments, the buffer andwash solution contains approximately 100 ppm benzalkonium chloride orProClin 300®.

In some embodiments, the buffer and wash solution can include asurfactant. The surfactant may be non-ionic, cationic, anionic orzwitterionic. Mixtures of surfactants may also be used. Exemplaryclasses of surfactants include alcohol ether sulfates, alcohol sulfates,alkanolamides, alkyl sulfonates, amine oxides, amphoteric surfactants,anionic surfactants, betaine derivatives, cationic surfactants,disulfonates, dodecylbenzene, sulfonic acid, ethoxylated alcohols,ethoxylated alkyl phenols, ethoxylated fatty acids, glycerol estershydrotropes, lauryl sulfates, mono and diglycerides, non-ionicsurfactants, phosphate esters, quaternary surfactants, and sorbitanderivatives. Additional examples of surfactants suitable for embodimentsof the buffer and wash solution are disclosed in co-pending U.S. patentapplication Ser. No. 13/526,164 filed on Jun. 18, 2012, the disclosureof which is incorporated herein by reference in its entirety.

In some embodiments, the buffer and wash solution further includes anacid to adjust the pH. The acid can be any acid traditionally used toadjust the pH of a solution. For example, acetic acid, nitric acid,hydrochloric acid, phosphoric acid, formic acid, sulfuric acid, orcitric acid can be used. If necessary, a base can be added as well toreduce the pH to the desired level.

The buffer and wash solution can have a pH of from approximately 5 toapproximately 9 (e.g., from approximately 5.5 to 8.5, from approximately5.5 to 8, from approximately 5.5 to 7, from approximately 5.5 to 6, fromapproximately 5.8 to 7.2, from approximately 6.8 to 7.2, a pH ofapproximately 6.0, a pH of approximately 6.5, or a pH of approximately7).

One way to make the buffer and wash solution, is to add distilled ordeionized water to a mixing vessel to less than 100% (e.g.,approximately 90%) of the final desired volume. Calculated amounts of anantimicrobial agent (e.g., ProClin 300®), an acid (e.g., acetic acid),and a buffering agent (e.g., HEPES) can then be added to the water.Further water can be added to bring the solution to its final desiredvolume. The mixture can be mixed with a magnetic stir plate/stir barand/or an impeller for a minimum of about 30 minutes. Other ways toprepare the solution can be used. After mixing, a pH reading can becarried out on an aliquot of the wash solution using a pH meter (e.g., aMettler pH meter). In some embodiments, if the pH is not within adesired range, then further acid can be added to the wash solution untilthe desired pH is attained.

Finally, the buffer and wash solution can be filtered through a 0.45 μmfilter to remove any particulates before bottling. In some embodiments,a finer filter can be used. For example, a 0.1 to one μm filter (e.g., a0.2 μm filter, a 0.4 μm filter, a 0.8 μm filter) can be used to removeany microorganisms and/or particulates in the wash solution. In someembodiments, the buffer and wash solution can be stored in a five-literbottle. The pH of the final product can be measured, if desired.

Systems for Analyzing Samples

FIG. 2 shows an analysis system 59 for analyzing samples such as bodyfluids. As discussed in U.S. Patent Application Nos. US2009/0269799 andUS2011/0070606, and in U.S. patent application Ser. No. 12/943,687,systems for analyzing fluid samples can include subsystems andcomponents to inspect body fluids such as blood, cerebrospinal fluid,and lymph, or other fluids, e.g., that contain cells. Components caninclude a chassis 61, a sample preparation system 21, a sample carriertransport system 63, one or more processing stations 65, a slide outputstation 69, a sample carrier labeler 71, and a control unit 67.

The chassis 61 can support the components of the analysis system 59. Insome implementations, the chassis 61 is in the form of a platform or atable onto which system components are secured.

The analysis system 59 can include one or more processing stations 65 toperform various processes. When analyzing a biological fluid, processingstations 65 can include a sample applicator, a sample preparationstation, and/or one or more imaging stations. Additionally, the analysissystem 59 can contain stations that do not have processing components topossibly reserve the location for future needs or uses. Processingstations 65 can be positioned in a straight direction with respect toone another, or alternatively the processing stations 65 can bepositioned in an arc or other shapes based on system and/or spacerequirements.

To transport the samples to each of the stations of the analysis system59, a sample carrier transport system 63 can have a translating memberhaving two or more carrier retaining devices attached used to move thesample carriers 55 to each of the processing stations 65.

Although the analysis system 59 shown in FIG. 2 has one sample carriertransport system 63, in some implementations, the analysis system 59 caninclude two or more sample carrier transport systems 63. For example ananalysis system 59 can include two or more sample carrier transportsystems 63 working in parallel.

Systems and methods for analyzing body fluids are disclosed, forexample, in U.S. Patent Application Publication Nos. 2011/0070606 and2012/0149050, the entire contents of each of which are incorporatedherein by reference. As one example, sample preparation system 21 can beused to deliver a sample to a sample carrier. In some implementations,for analysis systems that analyze fluid samples such as body fluids(e.g., blood, bone marrow, urine, semen, bile, breast milk,cerebrospinal fluid, lymph, gastric fluid, mucus, peritoneal fluid,sweat, tears, and/or saliva), one or more fluid samples can be providedto the analysis system 59 in one or more sample containers 39 (e.g.,test tubes) arranged in sample container magazines (e.g., test tuberacks). In such implementations, the sample preparation system can beconfigured to remove a small amount of the fluid sample from a samplecontainer 39 (e.g., a test tube) and apply the sample to a samplecarrier (e.g., a glass slide) using the sample applicator. Such sampleapplicators can be powered hydraulically or pneumatically using suctionto withdraw the fluid sample from the sample container 39 and then usingpressure to dispense the fluid into or onto the sample carrier. Thesample preparation system can further include systems to clean anysample handling devices to minimize cross-contamination of samples.

Depending on the type of samples analyzed by the analysis system 59,other types of sample applicators are possible. For example, if tissuesamples are analyzed, the sample applicator could include a mechanicaldevice to pick up the tissue, such as tweezers or forceps, and depositthe tissue sample evenly across the surface of a sample carrier.

Some sample types such as body fluids (e.g., blood, bone marrow, urine,semen, bile, breast milk, cerebrospinal fluid, gastric fluid, mucus,peritoneal fluid, lymph, sweat, tears, vomit, and/or saliva) can beanalyzed with a stain applied to permit certain types of visualinspection. In such analysis systems, a sample preparation station canbe provided to apply one or more fixative, stain, and/or rinse solutionsto the sample carried by the sample carrier.

To inspect or analyze the sample using the imaging station, a lightsource is generally included in the analysis system to illuminate thesample. Depending on the type of analysis to be conducted, the lightsource can include various types of visible light sources (e.g., lightemitting diodes, incandescent lights, fluorescent lights, and/or lasers)or non-visible light source (e.g., ultraviolet light and infrared lightsources). The positioning of the light source relative to the imagingstation can depend on the type of analysis conducted, as well as on thetype of sample carriers used. In some implementations, where samples arecarried on glass slides, an LED light source can be positioned below theglass slides to illuminate the sample.

The imaging station is electrically connected to the control unit 67 andcan be used to collect data from samples (e.g., can take an image of thesample to perform algorithms or analyses using the image). In someimplementations, the imaging station can use the image to performanalyses such as counting blood cells in a sample or to detect specificcells in the blood. As discussed above, in some implementations, thelight source can provide different forms of light so the imaging stationcan therefore include other types of detectors such as infrared lightdetectors or laser detectors used to measure certain properties (e.g.,dimensions) of the sample.

Once the analysis system has processed the sample at all of theprocessing stations 65, the slide output station 69 can disposition thesample, either discarding the sample or retaining the sample foradditional processing or future evaluation.

In such cases where it is desired to retain the sample and/or samplecarrier for additional processing or inspection, the sample carrierlabeler 71 (e.g., a printer device) can be used to provide sampleinformation to the sample and/or to the sample carrier. For analysissystems that analyze a patient's body fluids, the patient's informationcan be printed onto the sample carrier.

The control unit 67 can be electrically connected to the variouscomponents of the analysis system to control the operations of thecomponents, such as controlling the sample preparation system 21, samplecarrier transport system 63, the light source, the imaging system, andthe sample carrier labeler 71.

Providing a Sample to a Sample Applicator

FIG. 3 shows a flow chart describing an embodiment of a method andsequence of removing a sample from a sample container (e.g., a testtube) and providing the sample to a sample applicator system of a sampleanalysis system (e.g., a blood analysis system).

In this example, the sample preparation system includes one or moresample containers (e.g., test tubes) supported by a sample containermagazine (e.g., test tube rack), a sample container carrier, anextraction device (e.g., an extraction needle), a sample vessel tocontain and transport the sample through the sample preparation system,a wash cup for the extraction device, a modifying station, a sampleapplicator, a sample applicator wash cup, and a sample vessel washsystem to clean the sample vessel after the fluid sample has beenprovided to the sample applicator.

As shown in FIG. 3, to begin processing a first sample, a samplecontainer (e.g., a test tube) is removed from the sample containermagazine (step 73) (e.g., test tube rack) by a sample container carrier.As discussed above, the sample container carrier can include a roboticdevice (e.g., a robotic arm) that can articulate to grab or pick up thesample container and remove it from the sample container magazine.

The sample container carrier then places the sample container inposition for sample extraction (step 75) and a sample is extracted fromthe sample container (step 77). In some implementations, this includestranslating the sample container to be in line with an extractor needleand rotating the sample container such that it is positioned above theextractor needle, with a sample container cap (e.g., penetrable cap)pointed downward towards the extractor needle. In such implementations,the extractor needle is inserted through the sample container cap, whichcan be made of a rubber or plastic material, and into the samplecontainer far enough to penetrate through the cap to remove a liquidsample. When the sample is extracted with the sample carrier upsidedown, it can be more likely that a sample will be successfully extractedfrom the sample carrier, then when a sample is extracted with the samplecontainer positioned upright. When a sample is extracted with the samplecontainer positioned upright, a certain volume of fluid is typicallyrequired in the sample container to ensure that the extraction needlecan reach the fluid surface level during extraction. However, in otherimplementations, the sample can be extracted while the sample containeris in the upright position using methods to verify that a sample isproperly extracted.

Using either method of handling the sample containers and extracting asample, once the sample is extracted from the sample container, thesample container carrier can place the sample carrier back in the samplecontainer magazine (step 79). In some cases, the sample containers areplaced into the same position in the sample container magazine fromwhich they were removed prior to extraction of the sample. In othercases, the sample container can be placed in a different position withinthe sample container magazine, or alternatively the sample container canbe placed into a different sample container magazine than the samplecontainer magazine from which it was removed.

With the sample removed from the sample container, the sample can beplaced into a sample vessel using the extractor needle (step 81). Asdiscussed above, in some implementations, the extractor needle canoperate to handle the sample by using a hydraulic fluid, e.g., a bufferfluid, system to withdraw the sample and suspend it in the needle whilethe needle is in motion. In such implementations, the buffer fluid iscontrolled to move within the tubing and/or extractor needle just enoughto dispense the suspended sample into the sample vessel withoutdispensing the buffer fluid.

Once the sample is dispensed into the sample vessel, the sample vesselcan be moved away from the position where it receives the sample (e.g.,the sample extraction position) using a movement mechanism such as atrack (e.g., a pneumatic powered track, or a translating track andleadscrew device).

Once the sample vessel travels away from the sample extraction position,the extraction needle can be cleaned (step 83). As discussed above, insome implementations, the extraction needle can be inserted into a washcup and a portion of the buffer fluid can be expelled from the needle towash the inner surface and then the outer surface of the extractionneedle.

In some implementations, the sample vessel can be moved to amodification station prior to the sample being withdrawn from the samplevessel by a sample applicator (step 87). In such implementations, amodifying station can be positioned at a location along the trackbetween the extractor needle and the sample applicator. In some cases,the sample modification can include adding a diluent fluid to thesample. In such cases, a modification conduit (e.g., a fluid nozzle, aneedle, a syringe tip, a pipette tip, and/or a tubing portion) connectedto a modifying fluid reservoir is positioned along the track such thatthe sample vessel can stop under the modification conduit and a portionof the modifying fluid (e.g., diluent fluid) can be applied to thesample in the sample vessel (step 89).

Where the modification station includes preparing a sample using adiluent, diluents may include salt solutions or protein solutions. Saltsolutions range from “physiological saline” (0.9 N), to complex mixturesof salts, to the commercial preparation Plasmalyte that simulatesvirtually all the salts found in human blood serum. Protein solutionscan range from simple solutions of bovine albumin to Plasmanate®, acommercial preparation with selected human plasma proteins. Suchpreparations can vary in protein concentrations, buffers, pH,osmolarity, osmalality, buffering capacity, and additives of varioustypes. Synthetic or “substitute” versions of these solutions may also beusable, including Ficoll® or Dextran or other polysaccharides. Othersubstitutes may be used. An example of a diluent is Plasmalyte plusPlasmanate® in the proportion of 4:1 (Plasmalyte:Plasmanate®). Anotherexample of a diluent is 5% albumin. When preparing samples from wholeblood, a dilution of 2 parts blood to 1 part diluent can be used, wherethe diluent is a physiologically compatible solution, but a range ofdilution from 0:1 (no dilution) to 10:1 (diluent:blood) can be used inalternate embodiments.

In some implementations, the sample is not subjected to any modificationprior to being provided to the sample application, and thus the samplevessel and sample can be moved along the track from the sampleextraction position directly to the applicator position (step 91).

Once the sample vessel and sample is moved to the sample applicator, thesample applicator can withdraw the sample from the sample vessel (step93). The sample applicator can include an application conduit (e.g., afluid nozzle, a needle, and/or a tubing portion) connected to apneumatic or hydraulic fluid, e.g., buffer fluid, system, similar to thepneumatic or hydraulic system used with the extractor needle. To removethe sample from the sample vessel, vacuum can be applied to thehydraulic, e.g., buffer, fluid to generate low pressure in the tip ofthe application conduit. Such pressure can withdraw the sample into theapplication conduit such that an air gap between the buffer fluid andthe withdrawn sample fluid is generated. In some cases, it can beadvantageous to remove the entire fluid sample from the sample vessel.

After all or a portion of the sample is removed from the sample vessel,the sample vessel can be removed from the applicator position. In somecases, after the sample has been removed, the sample vessel istranslated away from the sample applicator back toward the sampleextraction position (e.g., at the extraction needle). In other cases,the track can extend beyond the location of the sample applicator, sothe sample vessel can move beyond the applicator position beforereturning back to the sample extraction position.

With the sample vessel no longer in the sample application position, thesample applicator can apply the sample to a surface such as a samplecarrier (step 95) (e.g., a cup, a flat plate, or a glass slide) so thatthe sample can be processed in another system (e.g., analyzed in ananalysis system). In some implementations, the sample applicator can beconnected to an articulating device that allows the sample applicator tomove to apply the sample to the surface. Typically, during applicationof a sample, the sample applicator dispenses substantially the entiresample contained in the sample applicator. Depending on the type ofsample and the requirements of the system in which the sample will beused, various application patterns (e.g., a boustrophedon pattern, araster pattern, a continuous spiral pattern, a pattern of multipleconcentric circles, and/or a pattern of multiple parallel lines) can beapplied to the application surface. Alternatively or additionally, thesample applicator can remain stationary and the sample carrier surfacecan be moved relative to the sample applicator to apply the appropriatepattern of sample.

In some cases, the sample applicator does not dispense all of the fluidsample volume when applying the sample. In such cases, some portion ofthe fluid sample can be retained in the sample applicator and/or aresidual amount of the fluid sample can accumulate on the outer surfaceor edge of the application conduit of the sample applicator. To avoidany cross contamination of samples, the application conduit of thesample applicator can be cleaned after applying the sample (step 101).Similar to the extractor needle, the application conduit of the sampleapplicator can be cleaned by inserting a distal portion of theapplication conduit into a wash cup and expelling a portion of thehydraulic, e.g., buffer fluid out of the application conduit into thewash cup that is shaped having a curved bottom to direct the flowexiting the application conduit up and over the outer surface of theapplication conduit to wash any fluid sample portion from both theinside surface and the outside surface of the application conduit.

Prior to returning to the sample extraction position, the sample vesselcan be moved to a sample vessel wash system to clean the sample vessel(step 97). In some implementations, as discussed above, the samplevessel wash system can include a nozzle to dispense a cleaning fluid toflush residual sample fluid from the sample vessel. The sample vesselwash system can also include a vacuum device to remove the cleaningfluid from the sample vessel, leaving the mix up cleaned. In otherimplementations, the sample vessel can include a drain device to disposeof the cleaning fluid provided to the sample vessel.

Once the sample vessel is cleaned, it can be moved along the track andreturned to the sample extraction position (e.g., under the extractorneedle) to receive a next sample obtained from a sample container (step99). In some implementations, the next sample can be obtained from anext sample container. However, alternatively, in other implementations,multiple sample aliquots (e.g., 2, 3, 4, 5, 10, or more sample aliquots)can be removed from the same sample container.

Although FIG. 3 shows one example of the sample preparation systemutilizing certain steps performed in a certain order to provide a sampleto a sample applicator, in other embodiments, the sample preparationsystem can include more or fewer steps, or some of the steps can beperformed in different orders. For example, although FIG. 3 shows step101 (cleanse sample applicator) as being the last step in the process,in other embodiments, the sample applicator can be cleaned before someof the other steps, e.g., before step 99 (translate sample vessel backto sample extraction position).

Example of a Sample Preparation System

FIG. 4 shows an example of a sample preparation system 21 used in abiological fluid analysis system. The sample preparation system 21removes a fluid sample such as a body fluid (e.g., blood, bone marrow,urine, semen, bile, breast milk, cerebrospinal fluid, gastric fluid,mucus, peritoneal fluid, sweat, tears, and/or saliva) from samplecontainers 39 (e.g., test tubes), and provides the sample to a sampleapplicator 31 of the biological fluid analysis system. The samplepreparation system 21 can include a sample container gripping device103, an information reading device 105 (e.g., a barcode reader), aninverting mechanism 47, a fluid extraction device 109, a sample vessel35, a sample vessel movement mechanism 33, a sample vessel wash system37, a sample modification system 29 (e.g., a diluent system), a sampleapplicator 31, an extraction needle wash cup 27, a sample applicatorwash cup 28, a fluid reservoir 26 a, a fluid pump 26 b, and a fluidsystem controller 26 c.

One or more sample containers 39 used in the sample preparation system21 can be in the form of test tubes having container closures 41 (e.g.,test tube caps) to contain the samples when the test tubes are moved oragitated. The test tube caps are typically made of a plastic material ora rubber material so that they can have the ability to re-seal puncturescreated by a needle (e.g., extraction needle 25). The test tubes aregenerally provided to the sample preparation system 21 in a samplecontainer magazine 45 (e.g., a test tube rack) for simplified storageand transport of multiple sample containers 39.

In some implementations, the sample containers 39 are test tubes and thetest tubes can have sample information 43 (e.g., machine-readableinformation such as a barcode) printed onto their outside surface. Insome cases, sample information 43 can include the type of sample, theorigin of the sample, the time and/or date when the sample was obtained.

The gripping device 103 can be used to remove a test tube 39 from thetest tube rack 45. As shown, in some implementations, the grippingdevice 103 can include two, three, or more finger members 102 that useradial motion to articulate inward and outward to temporarily retain atest tube 39. In some cases the finger members can move inward to gripthe test tube 39 by a bottom lip of the test tube cap 41 such that thegripping device 103 provides a lifting force to the test tube cap 41instead of a clamping force to the test tube 39, which could potentiallydamage the test tube 39. Such a gripping device 103 also allows forretrieving a particular test tube 39 in a limited space envelope,without disturbing surrounding test tubes 39 that may be present. Insome cases, radial motion used to articulate the finger members inwardand outward can be achieved by a gear system, a cam system, or byelectromechanical systems.

In other implementations, other mechanisms could be used to grip thetest tube 39 and remove it from the test tube rack 45. For example,other mechanical systems or magnetic systems can be used. In suchmagnetic systems, the test tube 39 and/or test tube caps 41 couldinclude magnetic portions and the gripping device 103 could include anelectromagnetic device that could be activated to magnetically fasten tothe test tube 39 and/or test tube cap 41 and pick it up.

As shown in FIG. 5A, in some implementations, the gripping device 103can remove the test tube 39 from the test tube rack 45 and also rotatethe test tube 39 about its longitudinal axis using electromagneticand/or mechanicals systems (e.g., electric motors, servos, gears, and/orcams) to pass the test tube 39 by an information reading device 105,such as a barcode reader. As discussed above and shown in FIG. 5A, insome implementations, the test tube 39 can have sample information 43(e.g., in the form of a barcode) printed on the outer surface of thetest tube 39, so as the test tube 39 is rotated in front of the barcodereader 105, the barcode reader 105 can read the barcode 43 to obtaininformation regarding the sample. In other implementations, the barcodereader 105 can be mounted on an articulating member so that the barcodereader 105 can move around a test tube 39 to read the barcode.

Alternatively, in some implementations, barcodes or othermachine-readable information can be applied to the test tube 39 atcertain locations relative to a point of reference on the test tube 39(e.g., at a certain angle with respect to the position of the test tube39) such that as the test tube is removed from the test tube rack 45,the gripping device 103 can be programmed to rotate the test tube 39 sothat a certain portion of the test tube 39 (e.g., the portion containingthe barcode) is in a position in front of the barcode reader 105.

In some implementations, the gripping device 103 can be mounted on atranslating track 117 (e.g., a linear actuator or an xyz robot) to movethe gripping device 103 from the test tube rack 45 where it can pick upa test tube 39, move the test tube 39 in front of the barcode reader105, and then provide the test tube 39 to the inverting mechanism 47.

FIG. 5B shows a gripping device 103 having a cap detector cover 104(shown in FIG. 5B to be semi-transparent to show the interiorcomponents) that is used to detect a test tube 39 and/or a test tube cap41 positioned on a test tube 39 in a test tube rack 45. The cap detectorcover 104 partially encloses multiple pinion gears 106 that rotate tomove two or more finger members 102 inward and outward radially to gripand release a test tube 39. The cap detector cover 104 is typically freeto translate in the longitudinal direction and is prevented fromrotating about its longitudinal axis relative to the pinion gears 106 byone or more rotation limiting devices 108 (e.g., shoulder screws) thatcan also act as bottom stops to provide a lower resting position for thecap detector cover 104. Typically, the force of gravity and the weightof the cap detector cover 104 are sufficient to cause the cap detectorcover 104 to rest at its lowest position along the shoulder screws 108.In some implementations, a deflection device 110 such as a spring orweight can be included to provide additional force to cause the capdetector cover 104 to be at rest at its lowest point.

As shown in FIG. 5B, a sensor 112, such as an optical sensor (e.g., anoptical isolator sensor) can be positioned above the cap detector cover104 so that when the gripping device 103 is lowered to a test tube 39,an elongated boss 114 of the cap detector cover 104 can contact the topof the test tube cap 41. Since the cap detector cover 104 is able tomove relative to the gripping device 103, when the cap detector cover104 is prevented from moving downward due to the presence of the testtube 39 or test tube cap 41, the rest of the gripping device 103continues to move downward and therefore the cap detector cover 104moves upward relative to the gripping device 103. The cap detector cover104 can continue to move upward relative to the rest of the grippingdevice 103 until the sensor 112 is tripped (e.g., when an outer portionof the cap detector cover 104 passes into a slot 112 a of the opticalisolator sensor 112 shown in FIG. 5B) and causes the gripping device 103to stop moving downward.

The sample preparation system 21 can use this method of articulatingdownward and detecting the position where the cap detector cover 104contacts a test tube 39 or a test tube cap 41 to determine whether atest tube 39 is present in the test tube rack 45, the size of the testtube 39 present in the test tube rack 45, and whether or not a test tube39 has a test tube cap 41 affixed on top by electronically storing andaccessing known ranges (e.g., positions) at which the gripping device103 should expect to contact a test tube 39 or a test tube cap 41.

For example, for sample preparation systems 21 that are configured toremove samples from test tubes 39 that are 75 mm or 100 mm long, if atest tube 39 is present in the test tube rack 45, the cap detector cover104 should make contact with the top of a test tube cap 41 and trip thesensor 112 at one of at least two positions (i.e., positions associatedwith where a test tube cap 41 affixed to the top of a test tube 39should be) located at distances from the bottom of a test tube rack, thedistances corresponding to a test tube height (e.g., 75 mm or 100 mm)plus the height of a test tube cap. If the cap detector cover 104 failsto trip the sensor 112 at either of these predicted two positions, thesample preparation system 21 can be alerted that a test tube 39 is notpresent in the intended test tube rack location.

In addition to detecting if a test tube 39 is present in the test tuberack 45, the gripping device 103 can also utilize the motion of the capdetector cover 104 to determine whether or not a test tube cap 41 hasbeen inadvertently omitted from, or has fallen off of, a test tube 39present in the test tube rack 45. Similar to storing predicted positionswhere the sample preparation system 21 should expect the cap detectorcover 104 to contact the top of a test tube cap 41 affixed on a testtube 39, indicating that a test tube 39 having a test tube cap 41 isproperly positioned in the test tube rack 45, the system can also storepositions (e.g., in a computer control system) where the cap detectorcover 104 could contact the top of a test tube 39 that does not have atest tube cap 41 affixed thereon, indicating that a test tube 39 ispositioned in the test tube rack 45, however the test tube 39 would nothave a test tube cap 41. Therefore during operation when the grippingdevice 103 translates downward to remove a test tube 39 from the testtube rack 45, the gripping device 103 can monitor the travel distance ofthe gripping device 103.

As the gripping device 103 moves downward, the sample preparation system21 can expect the sensor 112 to be tripped at a position that wouldindicate contact with a test tube cap 41 affixed onto the largest testtube (e.g., 100 mm test tube having a test tube cap). If the sensor 112is not tripped at that position, the gripping device 103 continues tomove downward and the sample preparation system 21 can expect the sensorto be tripped at a position that would indicate contact with the top ofthe largest test tube (e.g., 100 mm test tube) that does not have a testtube cap. If tripped at this position, it would indicate that a testtube 39 is positioned in the test tube rack 45, but that it does nothave a test tube cap 41 disposed thereon and the sample preparationsystem 21 should not remove and/or process that particular test tube. Ifthis occurs, the sample preparation system 21 conveys an error messageto an operator and/or logs the occurrence in an error log or equivalentrecord.

If the cap detector cover 104 fails to trip the sensor 112 at either ofthese positions associated with the top surface of a test tube cap 41positioned on a large test tube 39, or the top surface of the large testtube 39 itself, the gripping device 103 can continue to translatedownward to detect a smaller test tube (e.g., a 75 mm test tube).Similar to having predicted positions to detect a large test tube, thesample preparation system 21 can have predicted positions where itexpects the cap detector cover 104 to contact the top surface of a testtube cap 41 positioned on the smaller test tube 39, or the top surfaceof the smaller test tube itself 39. Similar to the large test tube 39,if the cap detector cover 104 does not contact the top surface of a testtube cap 41 affixed on a small test tube 39, the gripping device 103will continue to translate to determine if a small test tube 39 ispositioned in the test tube rack 45 without a test tube cap 41. If thesample preparation system 21 detects a small test tube 39 positioned inthe test tube rack 45 without a test tube cap 41 disposed thereon, thesample preparation system 21 can be directed to not remove thatparticular test tube 39 for processing.

If the cap detector cover 104 fails to trip the sensor 112 at any of theexpected positions, the sample preparation system 21 can determine thatno test tube 39 is present in that particular test tube rack positionand the sample preparation system 21 can alert an operator of the error,or alternatively log the error in an internal system, and move on to anext test tube 39 to be processed. Alternatively or additionally, insome implementations, a camera system can be used to verify properplacement of test tubes 39 within a test tube rack 45 during removal ofthe test tube 39 from the test tube rack 45 or during replacement of atest tube 39 to a test tube rack 45.

Once the sample preparation system 21 determines that the grippingdevice 103 is properly positioned above a test tube 39 having a testtube cap 41 to be processed, the finger members 102 can move inwardradially to grip the test tube 39 by the test tube cap 41. As shown inFIGS. 5B and 5C, the gripping device 103 includes a cam device 106, suchas a set of planetary gears, with each finger member 102 affixed at anoff-center position on each gear 106. The planetary gears 106 can bemounted on a gripping device rotating frame 116. In the illustratedexample shown in the cross-sectional view of FIG. 5D, the gear driverotating assembly can include a motor 118 having a shaft that isconnected to a gear drive rotating assembly including a coupling 120,pinion shaft 122 having a central gear 124 attached to its lower end,and a clutch mechanism 126 (e.g., a friction clutch) connected to thepinion shaft 122 and the rotating frame 116.

As the shaft of the motor 118 rotates, the gear drive rotating assemblycan also rotate. As the central gear 124 rotates due to the motorrotation, the outer planetary gears 106 rotate accordingly (shown indetail in FIG. 5C). As the planetary gears 106 rotate, the fingermembers 102 that are mounted at off-center positions along the planetarygears 106 move inward or outward radially. With the finger members 102moving inward radially, the motor 118 can continue to rotate until thefinger members 102 contact a test tube and cannot move further inward.FIG. 5D shows a cross-sectional view of the gripping device rotatingassembly.

As shown in FIG. 5D, the pinion shaft 122 that is connected to thecentral gear 124 is also connected to the friction clutch 126 such thatwhen a sufficient resistive force is applied to the rotation of thecentral gear 124 with respect to the planet gears 106, the frictionclutch 126 is engaged and the entire rotating frame 116 rotates aboutits central axis. Therefore, when the finger members 102 move inwardradially and contact a test tube 39, the friction clutch 126 engages andthe rotating assembly and test tube 39 begin rotating about theirlongitudinal axis (shown in FIG. 5A). As discussed in detail above, thisrotational motion of the test tube 39 can be used to rotate the testtube 39 in front of a machine to read information contained on the outersurface of the test tube (e.g., a barcode reader 105). Similar to theway the rotating frame 116 can rotate when the finger members 102 aremoved inward and reach a mechanical stop (e.g., the test tube), thefriction clutch 126 can also cause the rotating frame 116 to rotate inthe opposite direction when the finger members 102 have reached theirmost outward radial positions (e.g., after a test tube is released) andthe finger members come into contact with the rotating frame 116.Rotating the gripping device 103 in such a manner after it releases atest tube 39 can be performed in cases where it is desired for thefinger members 102 to be positioned in a certain orientation when thegripping device 103 is translated downward to grip a test tube 39 (e.g.,in cases where the test tubes can be packaged tightly together).

In addition to the benefits described above, generating the inward andoutward radial motion of the finger members 102 using the rotating gears106 allows for on-the-fly adjustments of the sample preparation system21 and gripping device 103 so that different types of test tubes 39,such as test tubes having different diameters (e.g., 13 mm and 16 mm) oreven sample carriers having different outer shapes (e.g., circular,square, triangular, or other shapes) can be removed from test tube racks45 without using multiple gripping devices 103.

Although the test tubes 39 have been described as being provided whiledisposed in a test tube rack 45, in some implementations, the samplepreparation system 21 can alternatively or additionally obtain a samplefrom a test tube 39 that is positioned in an accelerated responsereceptacle, such as a priority drawer or rack 45 a (as shown in FIG.5E). The priority drawer or rack 45 a can include one or more aperturesconfigured to receive a test tube 39. During operation of the samplepreparation system 21, an operator may have a sample that he/she wishesto prepare for inspection before the other samples disposed in testtubes 39 position in the test tube racks 45. The priority drawer 45 aprovides a location in which an operator can place such high prioritysamples contained in test tubes 39 to undergo accelerated preparationand analysis by the system.

Accordingly, the sample preparation system 21 can be configured suchthat when an operator places a test tube 39 in one of the apertures inthe priority drawer 45 a, the sample preparation system 21 can processthe test tubes 39 in the priority drawer 45 a before processing theother test tubes 39 in the test tube rack 45. If more than one test tube39 is present in the priority drawer 45 a, the sample preparation system21 can process all of the test tubes 39 present in the priority drawer45 a before continuing on to process samples contained in test tubes 39in the test tube rack 45. When processing samples contained in testtubes 39 positioned in the priority drawer 45 a, the griping device 103can typically be operated in the same manner discussed above withrespect to removing test tubes 39 from a test tube rack 45.

As shown in FIGS. 5A, 5F, 5G, and FIGS. 6-8, the inverting mechanism 47can be a device including two or more closure members 119 (e.g.,clamping jaws) that can clamp a test tube 39 to rotate the test tube 39about an axis which is perpendicular to the longitudinal axis of thetest tube 39 (e.g., to turn the test tube 39 upside down). In someimplementations, the clamping jaws 119 can utilize electromechanicaldevices such as an electric motor and leadscrews or servos to open andclose the clamping jaws 119. An electric motor can be used to rotate theinverting mechanism 47.

In some implementations, depending on the type of sample contained inthe test tube 39, it can be beneficial to agitate the sample containedin the test tube 39 (e.g., blood can be agitated to re-suspend the bloodcells or to mix non-homogenous samples). In such implementations, theinverting mechanism 47 can be used to fully invert test tube 39 multipletimes (e.g., 10 times) to achieve a desired level of agitation. In otherimplementations, the inverting mechanism 47 can be used to partiallyrotate the test tube 39 without fully inverting it (e.g., rocking thetest tube 30°-70° one or more times) to achieve a desired level ofagitation. Once the sample is ready for sample extraction (e.g., oncethe sample has been agitated, if required), the test tube 39 can berotated such that the test tube cap 41 is pointed downward so a sampleportion can be extracted.

Referring back to FIG. 4, the fluid extraction device 109 is a deviceused to remove a sample from the test tube 39 and can include anextraction needle 25 and an extraction needle rotating mechanism 121positioned under the inverting mechanism 47. In some implementations, asdiscussed above, the test tube 39 can include a test tube cap 41, whichthe extraction needle 25 can penetrate. In such implementations, theextraction needle 25 can be fluidly connected to a hydraulic, e.g.,buffer, fluid system connected to the fluid reservoir 26 a and the fluidpumps 26 b. Using the fluid pumps 26 b, the fluid system can generatemovement and pressure changes within the buffer fluid to apply suctionand withdraw the sample into the extraction needle 25.

In some implementations, the test tube 39 is rotated into an invertedposition (e.g., the test tube cap 41 is pointed downward); therefore theextraction needle 25 is inserted upward into the test tube 39 to extracta sample. By extracting the sample from the test tube 39 while the testtube 39 is inverted, the extraction needle 25 typically only needs topuncture and barely penetrate the test tube cap 41 to contact thesample. If the test tube 39 was upright and the extraction needle 25 wasinserted from the top, a minimum sample volume and height in the testtube 39 would typically be required to ensure that the extraction needle25 would be in contact with the sample when inserted.

In some implementations, a sample preparation system 21 can include atest tube stop 130 to temporarily secure the test tube 39 in thevertical direction so that when an extraction needle 25 is inserted intoand/or removed from the test tube cap 41, the test tube 39 remains in adesired position to help ensure that the extraction needle 25 canproperly penetrate into and be removed from a test tube cap 41. As shownin FIG. 5F, the test tube stop 130 can be mounted on a hinge assembly132 to move away from the test tube path as the test tube 39 is movedtowards the inverting mechanism 47 and when the test tube 39 is grippedand rotated by the inverting mechanism 47, the test tube stop 130 canrotate (e.g., using a ring gear and a pinion gear attached to anelectric motor) around the hinge 132 to secure the test tube 39.

The test tube stop 130 can include a rotating portion 130 a and a fixedportion 130 b. The rotating portion 130 a is provided to prevent thetest tube 39 from moving upward (e.g., when the extraction needle 25 isinserted into the test tube cap 41) and can be configured to movebetween a small test tube position (i.e., at rest position) and a largetest tube position (i.e., deflected position). The fixed portion 130 bcan include a recess sized large enough to allow the extraction needle25 to pass through without creating an obstruction and prevents the testtube 39 from moving downward (e.g., when the extraction needle 25 isremoved from the test tube cap 41).

The rotating portion 130 a includes a spring mechanism that allows therotating portion 130 a to automatically return to an “at rest” positionwhen released from a deflected position. During operation, when a testtube 39 is present in the inverting mechanism 47, the test tube stop 130can rotate to temporarily secure the test tube 39 during fluidextraction. As shown in FIG. 5F, when a shorter test tube (e.g., a 75 mmlong test tube) is being processed, the rotating portion 130 a canremain in its at rest position to properly secure the shorter test tube.

As shown in FIG. 5G, the test tube stop 130 can be used withoutmodification to also secure a longer test tube (e.g., a 100 mm long testtube). As shown, when a longer test tube 39 is present in the invertingmechanism 47 and the test tube stop 130 approaches the test tube 39, thelower member of the rotating portion 130 a, which can contact the top ofa smaller test tube when a smaller test tube is present in the invertingmechanism 47, can contact the side of the larger test tube 39 (shown inFIG. 5G). Due to the lower member contacting the side of the test tube39, the rotating member 130 b rotates downward as it approaches the testtube 39 so that an upper member of the rotating portion moves into aposition to secure the larger test tube 39 during penetration and exitof the extraction needle 25 within the test tube cap 41. After fluidextraction, the test tube stop 130 can move away from the test tube 39so that the test tube 39 can be rotated upright for removal from theinverting mechanism 47. As the test tube stop 130 moves away from thetest tube 39, the spring mechanism can cause the rotating portion 130 ato return to its at rest position so that it can possibly receive ashorter or longer test tube in subsequent processing.

In some implementations, since the extraction needle 25 extracts thesample while the extraction needle 25 is pointed upward, it wouldtypically be rotated to provide the sample to the sample vessel 35(e.g., to point downward). One or more of various types of mechanisms(e.g., electric motors, electromagnetic devices, pneumatic actuators,leadscrews, and/or cam devices) can be used to rotate and position theextraction needle 25.

As shown in FIG. 4 (and in greater detail in FIGS. 6-9), in someimplementations, an electric motor and leadscrew can be used totranslate the extraction needle 25 up and down while a cam mechanism canbe used to provide rotation. To provide translation, the extractionneedle 25 can include a non-rotating member 123 that acts as a leadscrewnut, translating along the leadscrew as it is rotated by the electricmotor. As shown in FIG. 6, to provide rotation, the extraction needlerotating mechanism 121 can include the non-rotating member 123 connectedto a rotating member 125 using a fastener at a pivot location 127. Therotating member 125 includes a pin 129 that is mounted at an off-centerposition with respect to a central axis of the pivot location 127 andmoves along a profiled slot 131 to rotate the extraction needle 25(e.g., 180° around a horizontal axis).

When the extraction needle 25 is at a most downward location (e.g., awayfrom the inverting mechanism 47) and is pointed downward (e.g., awayfrom the inverting mechanism 47), the pin 129 is in the slot 131 at aposition above the non-rotating member 123 (e.g., the pin 129 is closerto the inverting mechanism 47 than the non-rotating member 123). As theleadscrew rotates and translates the extraction needle 25 upward, thepin 129 moves along the slot, following the profile of the slot. Theslot 131 is shaped such that the motion of the pin 129 relative to thepivot location 127 causes the extraction needle 25 to rotate. At aparticular location along the slot 131 profile, the pin 129 momentarilystops moving upward and the center axis of the pivot passes the centeraxis of the pin 129 in the upward direction, allowing the continuedrelative motion of the pin 129 and the pivot to cause the extractionneedle 25 to continue to rotate upwards until the pivot location 127 isdirectly above the pin 129 and the extraction needle 25 is pointingupward.

With the extraction needle 25 pointed upward, the needle can betranslated upward to pierce the cap 41 and to be inserted into the testtube 39 to extract a sample. As shown in FIG. 7, once the extractionneedle 25 barely penetrates the test tube cap 41 and comes in contactwith the sample, a portion of the sample can be withdrawn into theextraction needle 25 using the pneumatic or hydraulic techniquesdescribed herein, including the use of an air bubble or gap between thesample and the hydraulic fluid, e.g., a buffer fluid. The amount ofsample withdrawn into the extraction needle 25 can vary depending on theanalysis system in which the sample will be used. In someimplementations, the extraction needle 25 can extract 10-50 microliters(e.g., 15, 20, 25, 30, 35, 40, or 45 microliters, e.g., 10 to 30 or 35microliters) of the sample from the test tube 39. Samples can be removedwhile the test tube 39 is inverted in this manner even if the test tube39 does not contain a minimum amount of sample that would be required ifthe extraction needle 25 were inserted from the top to contact thesample and the test tube were being held upright.

As shown in FIG. 8, once the sample portion is withdrawn and containedin the extraction needle 25, the extraction needle 25 can be removedfrom the test tube 39 by translating the extraction needle 25 downward.In some implementations, due to the design of the test tube cap 41 andthe material it is made of, as the extraction needle 25 is removed fromthe test tube 39, the test tube cap 41 is able to automatically sealpunctures created by the extraction needle 25. As discussed above, asthe extraction needle 25 translates downward a cam device (e.g., theoff-center mounted pin 129 moving in the slot 131) causes the extractionneedle 25 to rotate 180° such that the extraction needle 25 containingthe sample portion is pointed downward.

As shown in FIG. 8, in some implementations, the downward pointedextraction needle 25 can be moved further downward to dispense thesample portion extracted from the test tube 39 into a sample vessel 35.In such implementations, the sample vessel 35 can be moved along thetrack 49 to be positioned under the extraction needle 25 (e.g., thesample extraction position) to receive the sample portion. To dispensethe sample into the sample vessel 35, pressure can be applied to thefluid system using the fluid pump 26 b such that the buffer fluid canmove in the extraction needle 25 and attached tubing to move the airbubble between the buffer fluid and the sample, and thus forcing thesample from the extraction needle 25 into the sample vessel 35. In someimplementations, all of the sample portion contained in the extractionneedle 25 can be dispensed into the sample vessel 35. In some cases, theamount dispensed can be between 10-50 microliters (e.g., 10 to 30microliters, e.g., 10, 15, 20, 25, 30, 35, 40, or 45 microliters).

Once the sample portion is dispensed into the sample vessel 35, theextraction needle 25 can move upward to prevent any alignment problems(e.g., so that the sample vessel 35 can move along the track 49 withoutinterference from the extraction needle 25).

In some implementations, instead of the gripping device 103 removing asample container 39, such as a test tube, from a test tube rack 45 forprocessing a fluid contained therein, an operator can manually provide atest tube 39 to a fluid analysis system so that the analysis system canprocess the fluid contained therein. In such implementations, the fluidanalysis system can include a door or opening that can open to receivethe test tube and fluid. FIG. 5H shows an open mode port aspirator 140in a stowed position that can be used to extract such samples from testtubes and provide the samples to the sample preparation system forprocessing. As shown in FIG. 5H, the open mode port aspirator 140 can bemounted to an xyz robot of the gripping device 103. The open mode portaspirator 140 can include an aspirator probe 142 having a seal 144(e.g., tapered conical seal) and can be connected to deploymentmechanism 146 (e.g., a four-bar linkage) to deploy the aspirator probe142 between a stowed position (shown in FIG. 5H) and a deployed position(shown in FIG. SI).

As shown in FIGS. 5H and 5I, the deployment mechanism 146 can include adevice to secure the aspirator probe in either the stowed position orthe deployed position, such as a two-position spring 148 connected tothe deployment mechanism 146. As shown in FIG. 5J, in addition to thefour-bar linkage that deploys the aspirator probe 142 outward radially,the deployment mechanism 146 can also move the aspirator probe 142 alonga semi-circular path (e.g., to swing the aspirator probe 142 along anarcuate path). To move the aspirator probe 142 along the arcuate path,the deployment mechanism 146 can include a hinge 150 that can beoperated by an arm of the xyz robot, which moves the gripping device103. The hinge 150 can further include a magnet set 152 to furthersecure the open mode port aspirator 140 and keep the hinge 150 closedwhile in a stowed position.

Once the aspirator probe 142 is in a fully deployed position (e.g.,positioned in an opening of the fluid analysis system), an operator canposition an uncapped test tube having a sample disposed therein aroundthe aspirator probe 142. In some cases, the test tube can positioned sothat a top surface of the test tube is seated along the tapered conicalseal 144 to prevent spilling of the sample contained therein duringaspiration. In some cases, the aspirator probe 142 can be configured toreach the bottom of most standard test tubes to help ensure that asample is removed from a test tube, even when the test tube containslittle fluid. However, full seating of the test tube along the taperedconical seal 144 is typically not required for operation. Similar to theother fluid handling devices of the sample preparation system (e.g., thefluid extraction device and sample applicator) the aspirator probe 142can be connected to a buffer fluid system used to withdraw the fluidfrom the test tube into the aspirator probe 142. Once the sample iswithdrawn into the aspirator probe 142, the operator can remove the testtube from the aspirator probe area. With the sample withdrawn and thetest tube removed, the xyz robot can translate the aspirator probe 142to a position above a sample vessel so that the sample can be dispensedinto the sample vessel for processing. Once in the sample vessel, thesubsequent processing of the sample is generally the same as if thesample were withdrawn using the extraction device.

Once the sample is dispensed from the aspirator probe 142, the xyz robotcan translate the aspirator probe to an aspirator probe wash station 142a (shown in FIG. 5J). In some implementations, the aspirator probe washstation 142 a can be a device in which the entire aspirator probe 142can be inserted and sealed using the tapered conical seal 144. A reasonfor having a separate aspirator probe washing station (i.e., as opposedto using the extraction needle wash station) is because since theaspirator probe can be inserted into test tubes containing a wide rangeof fluid levels, it is generally uncertain what portion of the outersurface of the aspirator probe 142 must be cleaned, and therefore usinga wash station that can clean the entire outer surface of the aspiratorprobe 142 can reduce contamination of subsequent samples.

Once the aspirator probe 142 is cleaned, the deployment mechanism 146can move the aspirator probe 142 back to a stowed position where it canremain until it is deployed for subsequent open mode port processing.

FIG. 16A shows another implementation of an open mode port aspirator240. In FIG. 16A, aspirator 240 is shown in an extended position suchthat a test tube or other sample container can be positioned inproximity to the aspirator, and the sample within the container can bedrawn into the aspirator. Aspirator 240 is mounted to an xyz robot ofgripping device 103, and includes an aspirator probe 242 having a seal244 (e.g., tapered conical seal). Aspirator 240 is connected todeployment mechanism 246 with a five-bar linkage to deploy the aspiratorprobe 142 between a stowed position and the deployed position shown inFIG. 16A.

In addition to the five-bar linkage that deploys the aspirator probe 242outward radially, deployment mechanism 246 can also move the aspiratorprobe 242 along a semi-circular path (e.g., to swing the aspirator probe242 along an arcuate path). To move the aspirator probe 242 along thearcuate path, the deployment mechanism 246 can include a hinge 250 thatcan be operated by an arm of the xyz robot, which moves the grippingdevice 103. The hinge 250 can further include a magnet set (not shown inFIG. 16A) to further secure the open mode port aspirator 240 and keepthe hinge 250 closed while in a stowed position.

Within the five-bar linkage, lower links 264 and 265 are connected byguide block 262, which keeps links 264 and 265 aligned with one another.Upper links 266 and 267 are directly connected and constitute the othermembers of the five-bar linkage. To move aspirator probe 242 into adeployed position, the xyz robot translates gripping device 103 so thataspirator probe 242 is inserted into an opening in U-shaped pivot block300, which is supported by paddle 302 and hood cover 304 as shown inFIG. 16B. FIG. 16C is a perspective view of a blood analyzer 3000, andshows the positions of paddle 302 and hood cover 304 in greater detail.As seal 244 comes into contact with the surface of pivot block 300, seal244 and aspirator probe 242 rotate outward within the opening so thataspirator probe 242 is in the deployed position shown in FIG. 16A.Individual links in the five-bar linkage move passively with respect toone another as aspirator probe 242 rotates. When the xyz robottranslates gripping device 103 upward in FIG. 16A, aspirator probe 242rotates inward toward a retracted position. The inward rotation of probe242 is halted by external spring 260 when probe 242 is fully retracted.Referring again to FIG. 16C, recess 3010 is contiguous with the openingin pivot block 300 so that aspirator probe 242 can be fully deployed andretracted, as shown in FIG. 16A.

With aspirator probe 242 in a fully deployed position (e.g., positionedin an opening of the fluid analysis system), an operator can position anuncapped test tube having a sample disposed therein around the aspiratorprobe 242. In some cases, the aspirator probe 242 can be configured toreach the bottom of most standard test tubes to help ensure that asample is removed from a test tube, even when the test tube containslittle fluid. Similar to the other fluid handling devices of the samplepreparation system (e.g., the fluid extraction device and sampleapplicator) the aspirator probe 242 can be connected to a buffer fluidsystem used to withdraw the fluid from the test tube into the aspiratorprobe 242. Once the sample is withdrawn into the aspirator probe 242,the operator can remove the test tube from the aspirator probe area.With the sample withdrawn and the test tube removed, the xyz robot cantranslate the aspirator probe 242 to a position above a sample vessel sothat the sample can be dispensed into the sample vessel for processing.

Once the sample is dispensed from the aspirator probe 242, the xyz robotcan translate the aspirator probe to an aspirator probe wash station andaspirator probe 242 can be cleaned as disclosed above. After cleaning,aspirator probe 242 can be moved by deployment mechanism 246 to a stowedposition where it can remain until it is deployed for subsequent openmode port processing.

Referring back to FIG. 4, the sample vessel 35 is a vessel having anarcuate inner surface (e.g., spherical, elliptical, or similar shapedsurface) used to carry (e.g., contain or support) a sample (e.g., afluid sample) as it is transported to various components of the system(e.g., under the extraction needle 25, under the diluent needle, underthe sample applicator 31). In some implementations, the sample vessel 35can contain a volume of approximately 10 microliters to 100 microliters(e.g., 70 microliters). As discussed above, in some implementations, thesample vessel 35 is typically made of an inert, smooth, non-porousmaterial that can reduce wetting of fluids (e.g., sample fluids)contained the sample vessel 35 so that the fluids are expected to flowmore easily to the bottom of the sample vessel 35. In someimplementations, the sample vessel 35 can be formed of variousmaterials, such as different types of plastics (e.g., PTFE,acetalhomopolymer, acetalcopolymer, acrylic, Ultem®, Teflon®, Delrin®,or Noryl®), glasses, and/or metal materials.

The sample vessel wash system 37 can include a fluid delivery system toprovide a wash fluid (e.g., an embodiment of the combined buffer andwash fluid solution described herein) to the sample vessel 35 to cleanthe sample vessel 35 and to flush any residual sample fluids from thesample vessel 35. In some implementations, the sample vessel wash system37 can include a vacuum conduit used to remove any fluids dispensed intothe sample vessel 35 by the fluid delivery system. As shown in FIG. 13,in some implementations, the sample vessel wash system 37 can be rigidlymounted and the sample vessel 35 can have an additional translatingdevice to move the sample vessel 35 to the sample vessel wash system 37for cleaning. In other implementations, the sample vessel wash system 37can be mounted on a translating device so that the sample vessel washsystem 37 can be moved to contact and clean the sample vessel 35. Asshown in FIG. 13, the sample vessel wash system can be mounted betweenthe sample applicator 31 and the sample modification system 29 so thatas the sample applicator 21 applies a sample to a sample carrier 55, thesample vessel 35 can move along the track 49 towards the sampleextraction position and stop along the way to clean the wash cup 35 toprepare to receive a next sample. As shown in FIG. 14, after beingcleaned of residual sample fluids in a sample vessel wash system 37, thesample vessel 35 can be returned to the sample extraction position toreceive and carry a next sample.

To avoid contaminating various components within the system andjeopardizing the accuracy of blood analysis results, it is important toclean sample vessel 35 as thoroughly as possible between samples. FIG.17 shows another embodiment of a sample vessel 35 and wash system 37. InFIG. 17, sample vessel 35 includes a mixing cup 421 and a base 423. Washsystem 37 includes channels 403 and 405 extending through sample vessel35, and a cap 407 dimensioned to fit into the conical opening in cup421. After a sample has been withdrawn from cup 421, a small portion ofthe sample typically remains on the interior walls of cup 421. To cleanthe cup, a wash solution is deposited into cup 421 and the solution(with remnants of the sample) is partially drawn out through channel 405to rinse the interior cup surface. The, cap 407 is lowered into cup 421.The wash solution and sample remnants that remain within cup 421 arecompressed against the interior surface of cup 421. A portion of thisliquid is again drawn out through channel 405. The remaining portion isforced upward along the interior surface of cup 421 and spills over intobase 423 from which it is drawn out through channel 403. Cap 407 is thenwithdrawn from the opening in cup 421, leaving the interior surface ofthe cup clean.

Cup 421 can be formed from a variety of materials. In someimplementations, cup 421 is formed from a hard material such as quartz.The hardness of quartz (or other material) should be sufficiently greatsuch that the surface of cup 421 is not scratched by extraction syringesthat deposit or remove fluids from cup 421, and the absence of scratcheson the cup surface prevents formation of pockets of fluids at thesurface. Moreover, biological materials generally have low adherence toquartz surfaces, so that the interior surface of cup 421 can be easilycleaned according to the steps disclosed above. Without wishing to bebound by theory, it is believed that the high surface energy of quartzhelps to prevent the adherence of biological materials and solutions.Other materials with large surface energies can also be used to form cup421, including Teflon®, stainless steel, and PTFE.

Typically, cup 421 is a permanent component of a blood analyzer.However, in some implementations, cup 421 is a disposable component thatcan be discarded after one or more samples have been deposited therein.Disposable cups 421 can be formed of materials such as various plasticsto reduce costs.

Referring back to FIG. 4, the sample vessel movement mechanism 33 isprovided to move the sample vessel 35 to each of the various positionsassociated with each of the components of the system (e.g., under theextraction needle 25, under the modification system 29, and under thesample applicator 31). The sample vessel movement mechanism 33 caninclude a track 49 (e.g., a sliding track) on which the sample vessel 35can smoothly translate and be moved. The movement of the sample vessel35 on the track 49 can be controlled by various devices, such aselectromechanical devices (e.g., an electric motor connected to aleadscrew), an electromagnetic device, or a pneumatically poweredactuator. In some implementations, the track 49 can have the device usedto move and control the motion of the sample vessel 35 built into thetrack 49, such as a linear actuator (e.g., a pneumatic linear actuatoror an electromechanical linear actuator).

As shown in FIG. 4, in some implementations, a sample preparation system21 can include a sample modification system 29 (e.g., a diluent system)to modify the sample prior to the sample being provided to the analysissystem (e.g., prior to reaching the applicator position). A diluentsystem can include a diluent conduit 53 (e.g., a section of tubing, asyringe tip, a pipette, or a needle) connected to a fluid deliverysystem to provide a diluent fluid to the sample. In someimplementations, a diluent fluid for a blood sample can include saltsolutions (e.g., “physiological saline” or Plasmalyte™), proteinsolutions (e.g., bovine albumin, Plasmanate®) and/or synthetic solutions(e.g., Ficoll®, Dextran™, or other polysaccharides). As shown in FIG. 9,during use, a sample vessel 35 can be translated along the track 49 to aposition under the modification system 29 (e.g., to the diluentposition) to receive a portion of diluent. The amount of diluent fluiddispensed into a blood sample can vary based on the sample. In somecases, diluent fluid can be dispensed into a sample, e.g., a bloodsample, to achieve a diluent fluid to blood ratio ranging from 0:1 (nodilution) to 10:1 (diluent:blood). When analyzing whole blood, adilution of 2 parts blood to 1 part diluent can be used. In someimplementations where the sample is blood, 10 microliters to 150microliters (e.g., 25, 30, 40, 50, 75, 100, or 125 microliters) ofdiluent can be dispensed into the sample vessel 35 to mix with a volumeof sample of 10 to 35 microliters (e.g., 15, 20, 25, or 30 microliters).

Referring back to FIG. 4, the sample applicator 31 can include anapplication conduit 57 (e.g., a section of tubing, a syringe tip, apipette, or a needle) connected to a buffer fluid handling system.Similar to the extraction needle 25, the buffer fluid handling systemconnected to the application conduit 57 can be used to withdraw thesample from the sample vessel 35 into the application conduit 57 andthen to dispense the sample onto a sample carrier (e.g., a glass slide)of the analysis system. The sample applicator 31 can further include atranslating device 32 such that the application conduit 57 can move inmultiple directions when applying the sample.

As shown in FIG. 11, in some implementations, the sample applicator 31(e.g., the application conduit 57) can be translated downward andinserted into the sample vessel 35 to withdraw the sample contained inthe sample vessel 35. In some implementations, the application conduit57 removes the entire sample, or a substantial majority of the sample.In certain implementations, the application conduit 57 can withdraw asample having a specific volume of between 0.1-50 microliters (e.g.,0.08 microliters, 1.0 microliters, 30 microliters). Similar to theoperation of the extraction needle 25 extracting the sample using thefluid system, the application conduit 57 can withdraw the sample intothe application conduit 57 by changing the pressure of the buffer fluid(e.g., using a fluid pump 26 b to reduce the buffer fluid pressure tocreate a vacuum in the application conduit 57) to cause the sample toflow into the application conduit 57.

As shown in FIG. 12, once the sample is withdrawn into the applicationconduit 57, it can be dispensed onto a sample carrier 55 (e.g., a glassslide) as the sample vessel 35 moves along the track 49 away from thesample application position. In some implementations, the sample can bedispensed using the same hydraulic, e.g., buffer, fluid system used towithdraw the sample into the application conduit 57. In someimplementations, the application conduit 57 can be moved relative to theglass slide 55 to produce various patterns of the sample fluid onto theglass slide 55. Such patterns can include a serpentine or rasterpattern, a continuous spiral pattern, a pattern of multiple concentriccircles, and/or a pattern of multiple parallel lines. In some cases, ablood sample can be applied to the glass slide to form a monolayer of asample containing cells, such as a sample of blood (e.g., a layer ofcells approximately one cell thick). In some implementations, the heightof the sample layer applied can range from less than 1 micron to 10microns or more. The sample can be applied in one continuous flow or inmultiple flows that are spaced apart or are applied side-by-side or evencontacting each other.

Although samples can be dispensed at various flow rates based on thetype of sample and the desired sample pattern formed on the glass slide55, in some implementations, an application conduit 57 having an innerdiameter of 300 microns can provide a sample flow rate of 0.1microliters per second. More generally, the inner diameter ofapplication conduit 57 can be in a range from 200 microns to 650 micronsinclusive (e.g., between 200 microns and 400 microns, between 300microns and 400 microns, between 400 microns and 650 microns, between500 microns and 650 microns).

In some implementations, the entire sample is dispensed from theapplication conduit 57 onto the glass slide. In some implementations,the flow rate of the sample dispensed from the application conduit 57can be 0.1 microliters per second while the application conduit 57 ismoving at a speed of 30 millimeters per second over the glass slidesurface at a height of about 5 to 100 microns, e.g., 15 to 50, 10 to 15,20 to 40, or 5 to 15 microns, about 12 microns. In some implementations,when dispensing a sample of undiluted blood, the flow rate through theapplication conduit 57 can be approximately 0.04 microliters per second,e.g., 0.02 to 0.10, 0.02 to 0.05, or 0.03 to 0.04 microliters persecond, while the application conduit 57 is moving at a speed of about50 millimeters per second, e.g., 10 to 100, 20 to 80, 30 to 70millimeters per second, while the application conduit 57 is at a heightof 10, 12, 14, 15, 20, or 25 microns from the slide surface.

Referring back to FIG. 4, in some implementations, the samplepreparation system can include multiple wash cups (e.g., an extractionneedle wash cup 27 and a sample applicator wash cup 28) that can operatein substantially the same way. Although the following explanation isdirected towards one particular wash cup (e.g., the extraction needlewash cup 27) and references components and/or features of the extractionneedle wash cup 27, the structure and operation of other wash cups inthe system (e.g., the sample applicator wash cup 28) are substantiallythe same as described.

The extraction needle wash cup 27 can include an inner basin 27 a and anouter basin 27 b that substantially surrounds the inner basin 27 a. Theinner basin 27 a can be shaped (e.g., a substantially cylindrical and/orhalf spherical inner basin) to receive a fluid dispensing member portion(e.g., the extraction needle 25 and/or the application conduit 57) andcan be designed to be slightly larger (e.g., 25% to 100% larger) thanthe outer diameter of the member portion (e.g., the extraction needle 25or the application conduit 57) to be inserted into the wash cup 27 andcan have a substantially rounded bottom. The outer basin 27 b caninclude a fluid output device 27 c (e.g., a drain or fluid suctiondevice). The inner basin 27 a and the outer basin 27 b can be designedsuch that when a member is inserted into the wash cup 27, the bufferfluid can be dispensed from the member to wash the inner surface of themember. When the buffer fluid exits the member portion, the roundedshape of the inner basin 27 a can cause the fluid to continuously flowupwards along the outer surface of the member as the wash cup fills withthe fluid. The continuous flow of buffer fluid pumped using the pump 26b from the fluid reservoir 26 a through the member portion and directedby curved bottom surface of the wash cup 27 allows the inserted memberportion to flush residual fluid samples remaining on the inner surfaceand/or the outer surface while minimizing the likelihood ofcross-contamination of samples.

Cross-contamination can be minimized because none of the wash fluidpumped from the member portion during rinsing/cleaning typicallyre-enters the member. After the buffer fluid flows along the outersurface of the member, it can flow over an upper edge of the inner basin27 a and into the outer basin 27 b. The contaminated fluid in the outerbasin 27 b can be removed by the fluid output device 27 c of the outerbasin 27 b and disposed into a waste reservoir. After a certain amountof buffer fluid is dispensed from the member, the fluid system can stopthe flow of the buffer fluid and withdraw the buffer fluid back into themember to create an air pocket in the member such that the air pocketcan serve as a barrier between the buffer fluid and future fluidswithdrawn into the member (e.g., a next portion of sample).

In some embodiments, wash cup 27 (and wash cup 28) can be implemented asdescribed above for sample vessel 35. For example, wash cup 27 caninclude an insert with a high surface energy (e.g., formed of a materialsuch as quartz) supported by a base. Wash cup 27 can have a geometrysimilar to that shown in FIG. 17 for vessel 35, and can be automaticallycleaned in a similar manner.

As shown in FIG. 10, the extraction needle 25 can be translated down andinserted into the extraction needle wash cup 27 to be cleaned as thesample vessel 35 is translating along to track 49 away from the sampleapplication position. Similarly, as shown in FIG. 15, as the samplevessel moves along the track 49 away from sample applicator 21, theapplication conduit 57 can be inserted into the sample applicator washcup 28 and cleaned by dispensing buffer fluid from the applicationconduit 57 such that the buffer fluid washes the inner surface and outersurface of the application conduit 57. Once cleaned in a wash cup, thefluid handling devices (e.g., the extraction needle 25 and theapplication conduit can handle a next sample).

With all of the components of the sample preparation system 21 cleaned,a next sample from the same or a next test tube 39 can be prepared foranalysis

Other Embodiments

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

What is claimed is:
 1. A method of handling a sample, the methodcomprising: receiving a sample container containing a volume of asample; removing a sample from the sample container using an extractiondevice; dispensing the sample into a sample vessel with the extractiondevice; washing the extraction device by dispensing a fluid through theextraction device; moving the sample vessel containing the sample to asample application position; removing a portion of the sample from thesample vessel using a sample applicator; dispensing the sample portionfrom the sample applicator onto a sample carrier; rinsing the sampleapplicator by dispensing fluid through the sample applicator; andwashing the sample vessel to remove any residual sample.
 2. The methodof claim 1, wherein the extraction device is operated by a fluid system,and wherein fluid of the fluid system is separated from the sample by anair pocket within the extraction device.
 3. The method of claim 1,further comprising modifying the sample dispensed into the samplevessel.
 4. The method of claim 3, wherein modifying the sample comprisesadding a diluent fluid to the sample in the sample vessel.
 5. The methodof claim 1, wherein removing a sample from the sample containercomprises inserting a needle through a cap attached to a test tube. 6.The method of claim 1, wherein the extraction device and the sampleapplicator retain the sample when the fluid control system generates avacuum in the extraction device and the sample applicator.
 7. The methodof claim 1, wherein rinsing the extraction device and rinsing the sampleapplicator comprises inserting a portion of the extraction device and aportion of the sample applicator into respective receptacles havingcurved bottoms, such that fluid dispensed from the extraction device andthe sample applicator is directed along an outer surface of theextraction device and the sample applicator.
 8. The method of claim 1,wherein dispensing the sample portion from the sample applicatorcomprises dispensing the sample portion onto a glass slide.
 9. Themethod of claim 1, wherein the sample comprises a body fluid.
 10. Themethod of claim 9, wherein the body fluid is blood.